Journal: Biochimica et biophysica acta

Article Title: Cyclopeptide RA-V inhibits cell adhesion and invasion in both estrogen receptor positive and negative breast cancer cells via PI3K/AKT and NF-κB signaling pathways.

doi: 10.1016/j.bbamcr.2015.04.020

Figure Lengend Snippet: Fig. 1. (A) Chemical structure of cyclopeptide RA-V. (B–E) Data of microarray analysis. (B) The heatmap of differential expressed genes in RA-V treated and untreated control samples (n = 3). Hierarchical cluster analysis was carried out for all differential expressed genes (fold change ≥2 or ≤0.5; p value b 0.05) in RA-V-treated Hela cells, compared with the untreated control. Each column represented one sample. Red colored columns indicated up-regulated genes while green colored columns indicated down-regulated genes. (C) Hierarchical cluster analysis showed the differential expressions of tumor metastasis-related genes induced by RA-V. Each column represented one sample and each row represented one gene. The scale of color intensity was positively correlated to the fold change. (IGFBP6, insulin-like growth factor binding protein 6; ECM2, extracellular matrix protein 2; ICAM4, intercellular adhesion mol- ecule 4; TIMP4, tissue inhibitor of metalloproteinase 4; ICAM3, intercellular adhesion molecule 3; ICAM5, intercellular adhesion molecule 5; PLAU, urokinase-plasminogen activator; ITGA2B, integrin, alpha 2b; LAMA3, laminin, alpha 3; ALDH3A2, aldehyde dehydrogenase 3 family, member A2; TP53, tumor protein p53; LAMB3, laminin, beta 3; ALDH5A1, aldehyde dehydrogenase 5 family, member A1; ALDH3B1, aldehyde dehydrogenase 3 family, member B1; IGFLR1, IGF-like family receptor 1; COL1A1, collagen, type I, alpha 1; CFL2, cofilin 2; MMP15, matrix metallopeptidase 15; ID1, inhibitor of DNA binding 1; IGSF8, immunoglobulin superfamily, member 8; ID3, inhibitor of DNA binding 3; TIMP3, tissue inhibitor of metal- loproteinase 3; ALDH16A1, aldehyde dehydrogenase 16 family, member A1; MMP28, matrix metallopeptidase 28; IGSF3, immunoglobulin superfamily, member 3; LAMB2, laminin, beta 2; COL4A5, collagen, type IV, alpha 5; COL4A4, collagen, type IV, alpha 4; IGFL2, IGF-like family member 2; TNC, tenascin C; IGF2BP1, insulin-like growth factor 2 mRNA binding protein 1; LAMA2, laminin, alpha 2; MMP7, matrix metallopeptidase 7; MMP19, matrix metallopeptidase 19; ITGB8, integrin, beta 8; ITGAM, integrin, alpha M; LAMB1, laminin, beta 1; r, laminin, gamma 2). (D) RA-V induced cancer invasion and metastasis-associated GO categories. GO categories cover 3 domains, cellular component, molecular function and biological process. Genes with differential expressions were analyzed and subjected to GO analysis in accordance with the SBC Analysis System. GO categories (p value b 0.05; Hit ≥5) were considered to be significant. ‘Hits’ was considered as the number of the differential expressed genes according to each GO category. (E) Table showed the top 5 signaling pathways involved in Hela cells in response to RA-V. aThe number of the differential expressed genes in a pathway according to Biocarta and Kegg databases. bThe enrichment p value of the pathway as de- termined by R-package Fisher's Exact Test.

Article Snippet: The blots were incubated with primary antibodies, betaactin (Sigma, USA), MMP1, MMP2, MMP7, MMP9, uPA, TIMP-1, ER, ROCK1, pEGFR, EGFR (Abcam, USA), cofilin, p-cofilin, Cdc42, RhoA, TIMP-2, PI3K, pPI3K, AKT, pAKT, FAK, NF-κB, pNF-κB, IκB and pIκB (Cell Signalling, USA) overnight.

Techniques: Microarray, Control, Binding Assay, Protein-Protein interactions